A novel and high-efficient method for the preparation of heat-stable antifungal factor from Lysobacter enzymogenes by high-speed counter-current chromatography

Frontiers in Microbiology

Microbiology

Liangliang Dong¹  Jianhong Xu²  Yancun Zhao³  Fengquan Liu³  Weibo Sun³  Bao Tang³ 

[1] College of Plant Protection, Anhui Agricultural University, Hefei, China;Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing, China;Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, China;

关键词: heat-stable antifungal factor;    Lysobacter enzymogenes;    separation;    preparation;    high-speed counter-current chromatography;   

DOI  :  10.3389/fmicb.2023.1227244

 received in 2023-05-23, accepted in 2023-07-27,  发布年份 2023

来源: Frontiers

【 摘 要 】

Heat-stable antifungal factor (HSAF) produced by the biocontrol bacterium Lysobacter enzymogenes shows considerable antifungal activity and has broad application potential in the agricultural and medical fields. There is a great demand for pure HSAF compounds in academic or industrial studies. However, an efficient preparation method that produces a high yield and high purity of HSAF is lacking, limiting the development of HSAF as a new drug. In the present study, high-speed counter-current chromatography (HSCCC) combined with column chromatography was successfully developed for the separation and preparation of HSAF from the crude extract of L. enzymogenes OH11. The crude extract was obtained by macroporous resin adsorption and desorption, and the main impurities were partly removed by ultraviolet light (254 nm) and gel filtration (Sephadex LH-20). In the HSCCC procedure, the selected suitable two-phase solvent system (n-hexane/ethyl acetate/methanol/water = 3:5:4:5, v/v, the lower phase added with 0.1% TFA) with a flow rate of 2.0 mL/min and a sample loading size of 100 mg was optimized for the separation. As a result, a total of 42 mg HSAF with a purity of 97.6% and recovery of 91.7% was yielded in one separation. The structure elucidation based on HR-TOF-MS, 1H and 13C NMR, and antifungal activities revealed that the isolated compound was unambiguously identified as HSAF. These results are helpful for separating and producing HSAF at an industrial scale, and they further demonstrate that HSCCC is a useful tool for isolating bioactive constituents from beneficial microorganisms.

【 授权许可】

Unknown   
Copyright © 2023 Sun, Tang, Dong, Xu, Zhao and Liu.